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In Vitro Assessment of Nanoparticle Effects on Blood Coagulation

  • Timothy M. Potter
  • Jamie C. Rodriguez
  • Barry W. Neun
  • Anna N. Ilinskaya
  • Edward Cedrone
  • Marina A. DobrovolskaiaEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1682)

Abstract

Blood clotting is a complex process which involves both cellular and biochemical components. The key cellular players in the blood clotting process are thrombocytes or platelets. Other cells, including leukocytes and endothelial cells, contribute to clotting by expressing the so-called pro-coagulant activity (PCA) complex on their surface. The biochemical component of blood clotting is represented by the plasma coagulation cascade, which includes plasma proteins also known as coagulation factors. The coordinated interaction between platelets, leukocytes, endothelial cells, and plasma coagulation factors is necessary for maintaining hemostasis and for preventing excessive bleeding. Undesirable activation of all or some of these components may lead to pathological blood coagulation and life-threatening conditions such as consumptive coagulopathy or disseminated intravascular coagulation (DIC). In contrast, unintended inhibition of the coagulation pathways may lead to hemorrhage. Thrombogenicity is the property of a test material to induce blood coagulation by affecting one or more elements of the clotting process. Anticoagulant activity refers to the property of a test material to inhibit coagulation. The tendency to cause platelet aggregation, perturb plasma coagulation, and induce leukocyte PCA can serve as an in vitro measure of a nanomaterial’s likelihood to be pro- or anticoagulant in vivo. This chapter describes three procedures for in vitro analyses of platelet aggregation, plasma coagulation time, and activation of leukocyte PCA. Platelet aggregation and plasma coagulation procedures have been described earlier. The revision here includes updated details about nanoparticle sample preparation, selection of nanoparticle concentration for the in vitro study, and updated details about assay controls. The chapter is expanded to describe a method for the leukocyte PCA analysis and case studies demonstrating the performance of these in vitro assays.

Key words

Nanoparticles Thrombogenicity Platelet aggregation Platelet Blood Plasma coagulation Coagulation factors Leukocyte procoagulant activity Thrombosis Disseminated intravascular coagulation DIC 

Notes

Acknowledgment

This project has been funded in whole or in part with Federal funds from the National Cancer Institute, National Institutes of Health, under Contract No. HHSN261200800001E. The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the U.S. Government.

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Copyright information

© Springer Science+Business Media LLC 2018

Authors and Affiliations

  • Timothy M. Potter
    • 1
  • Jamie C. Rodriguez
    • 1
  • Barry W. Neun
    • 1
  • Anna N. Ilinskaya
    • 1
  • Edward Cedrone
    • 1
  • Marina A. Dobrovolskaia
    • 1
    Email author
  1. 1.Cancer Research Technology Program¸ Nanotechnology Characterization LaboratoryLeidos Biomedical Research, Inc., Frederick National Laboratory for Cancer ResearchFrederickUSA

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